Recombinant Human CD39L4/ENTPD5 Protein, CF Summary
Val21-His428 (major) & Ser46-His428 (minor), both with a C-terminal 6-His tag
Accession # O75356
Complete Your Research
Complete Your Experiment
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
|Formulation||Supplied as a 0.2 μm filtered solution in Tris, CaCl2, NaCl and Glycerol.|
|Shipping||The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.|
|Stability & Storage:||Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- Assay Buffer: 25 mM Tris, 5 mM CaCl2, pH 7.5
- Recombinant Human CD39L4/ENTPD5 (rhCD39L4) (Catalog # 5297-EN)
- Substrate: GDP (Sigma, Catalog # G-7127), 100 mM stock in deionized water
- Malachite Green Phosphate Detection Kit (Catalog # DY996)
- 96-well Clear Plate (Costar, Catalog # 92592)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute rhCD39L4 to 0.04 µg/mL in Assay Buffer.
- Prepare a standard curve from 1 M Phosphate Standard by adding 10 µL of the 1 M Phosphate Standard to 990 µL of Assay Buffer for a 10 mM stock. Continue by adding 10 µL of the 10 mM Phosphate stock to 990 µL of Assay Buffer for a 100 µM stock (this is the first dilution to use as a standard).
- Perform six additional one-half serial dilutions of the 100 µM Phosphate stock in Assay Buffer. The standard curve has a range of 0.039 to 2.5 nmol per well.
- Load 25 µL of 0.04 μg/mL rhCD39L4 and the standard curve into a plate. Include a Substrate Blank containing Assay Buffer.
- Dilute the Substrate to 100 µM in Assay Buffer.
- Add 25 µL of the 100 µM Substrate to all wells and mix well.
- Cover the plate with parafilm or a plate sealer and incubate at 37 °C for 20 minutes.
- Add 10 µL of the Malachite Green Reagent A to all wells. Mix and incubate for 10 minutes at room temperature.
- Add 10 µL of the Malachite Green Reagent B to all wells. Mix and incubate for 20 minutes at room temperature.
- Read plate at 620 nm (absorbance) in endpoint mode.
- Calculate specific activity:
Specific Activity (pmol/min/µg) =
|Phosphate released* (nmol) x (1000 pmol/nmol)|
|Incubation time (min) x amount of enzyme (µg)|
*Derived from the phosphate standard curve using linear or 4-parameter fitting and adjusted for Substrate Blank.Per Well:
- rhCD39L4: 0.001 µg
- Substrate: 35.7 µM
Ectonucleoside triphosphate diphosphohydrolase-5 (NTPDase-5) is a protein that can be either secreted or reside in the lumen of the endoplasmic reticulum. The enzyme displays a specificity for nucleoside-5'-diphosphates, preferentially hydrolyzing GDP, IDP, and UDP to produce nucleoside monophosphates and orthophosphate (1). The enzyme is most highly expressed in liver, kidney, prostate, testis, and colon. NTPDase-5 (ENTPD5) and proto-oncogene CPH (PCPH) are encoded by the same gene (2). Alterations in ENTPD5/PCPH expression and mutations of ENTPD5 are associated with the progression of testicular (3), laryngeal (4), and other cancers.
- Mulero, J.J. et al. (2000) Biochemistry. 39:12924.
- Paez, J.G.. et al. (2001) Int. J. Oncol. 19:1249.
- Regadera, J. et al. (2006) Int. J. Oncol. 28:595.
- Blanquez, M.J. et al. (2002) Mol. Carcinogen. 35:186.
Citation for Recombinant Human CD39L4/ENTPD5 Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
1 Citation: Showing 1 - 1
Blocking Antibodies Targeting the CD39/CD73 Immunosuppressive Pathway Unleash Immune Responses in Combination Cancer Therapies
Authors: I Perrot, HA Michaud, M Giraudon-P, S Augier, A Docquier, L Gros, R Courtois, C Déjou, D Jecko, O Becquart, H Rispaud-Bl, L Gauthier, B Rossi, S Chanteux, N Gourdin, B Amigues, A Roussel, A Bensussan, JF Eliaou, J Bastid, F Romagné, Y Morel, E Narni-Manc, E Vivier, C Paturel, N Bonnefoy
Cell Rep, 2019;27(8):2411-2425.e9.
Sample Types: Protein
Applications: ELISA Capture
No product specific FAQs exist for this product, however you mayView all Proteins and Enzyme FAQs
Phosphatase Activity Assays
Reviews for Recombinant Human CD39L4/ENTPD5 Protein, CF
There are currently no reviews for this product. Be the first to review Recombinant Human CD39L4/ENTPD5 Protein, CF and earn rewards!
Have you used Recombinant Human CD39L4/ENTPD5 Protein, CF?
Submit a review and receive an Amazon gift card.
$25/€18/£15/$25CAN/¥75 Yuan/¥1250 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image